A Alcaligenes strain ho-1 and its application

A technology of HO-1 and alcaligenes, which is applied in the direction of bacteria, biological water/sewage treatment, sustainable biological treatment, etc., can solve the problems of cumbersome operation, low organic load tolerance threshold, slow reactor start-up, etc., and achieve simplification Denitrification process flow, high-efficiency ammonia nitrogen removal capacity, and the effect of saving treatment space and cost

Active Publication Date: 2021-10-01
INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is the most widely used and mature denitrification technology so far. However, it has defects and limitations, mainly including: First, the environmental conditions of nitrification and denitrification bacteria are different, and they need to be carried out in two isolated reactors, or use Intermittent aerobic and anaerobic conditions will cause problems of high cost and cumbersome operation; secondly, nitrifying bacteria are chemoautotrophic bacteria, on the one hand, their growth and reproduction time is longer, which makes the reactor start-up slow, thereby increasing On the other hand, it has a low tolerance threshold for organic loads, is sensitive to environmental impacts, and is difficult to operate; in addition, denitrifying bacteria are heterotrophic bacteria that need organic matter, while nitrifying bacteria cannot tolerate organic matter, so in During sewage treatment, organic matter is usually removed before the nitrification process, and carbon sources are added during the denitrification stage, which makes the process operation unable to be effectively simplified

Method used

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  • A Alcaligenes strain ho-1 and its application
  • A Alcaligenes strain ho-1 and its application
  • A Alcaligenes strain ho-1 and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Identification of bacterial strain HO-1

[0027] The Alcaligenes sp. strain HO-1 of the present invention is isolated from a short-path nitrification reactor for wastewater treatment in pig farms.

[0028] Inoculate strain HO-1 on LB solid slant medium, cultivate at 30°C until obvious bacterial lawn appears, pick a small amount of bacterial lawn with a sterile inoculation loop and mix it in an EP tube filled with 200 μL sterile water, vortex Mix to make a bacterial suspension. Then put the EP tube into the boiling water bath and cook for 3 minutes, and put it on ice immediately after taking it out. Bacteria cells are prone to rupture after transient hot and cold alternation, and then release genomic DNA. The lysed bacterial suspension was boiled and centrifuged for a short time, and the supernatant was taken out, which could be used as a template for subsequent 16S rRNA gene PCR amplification.

[0029] The bacterial universal primer 27F / 1492R (27F: 5'-AGAG...

Embodiment 2

[0036] Embodiment 2. The growth and ammonia nitrogen removal of bacterial strain HO-1 with ammonia nitrogen as the only nitrogen source

[0037] Inoculate the HO-1 strain cryopreserved in glycerol into a test tube containing LB liquid medium, culture it in a shaker at 30°C and 160rpm for 24h to activate it, and then transfer 1% of the inoculum into a test tube containing 100mL LB liquid medium Cultivate in a 250mL Erlenmeyer flask at 30°C and 160rpm for 20h to be a seed culture solution.

[0038] The seed culture solution was centrifuged at 6000 rpm for 5 minutes to collect the cells, washed with sterile normal saline for 3 times, and then resuspended with an equal volume of sterile normal saline to prepare a seed suspension.

[0039] The above seed suspension was inoculated into a 500mL Erlenmeyer flask containing 200mL of HNM medium at an inoculation amount of 1.5%, and cultured at 30°C and 160rpm for 48h. Take out 3mL culture medium regularly, 1mL is used for measuring the...

Embodiment 3

[0046] Example 3. Detection of Nitrogen Products in the Denitrification Process of Bacterial Strain HO-1

[0047] Activate the bacterial strain HO-1 according to the method in Example 2, take 4 mL of seed culture solution, centrifuge at 6000 rpm for 5 minutes to collect the bacteria, wash the bacteria three times with sterile normal saline, and then resuspend with 8 mL of normal saline to prepare 2-fold diluted seeds Suspension.

[0048] Use a 2mL syringe to inoculate 1.2mL of the above seed suspension into a 500mL airtight bottle (not ventilated) containing 40mL nitrogen detection medium. Shaker culture. After 4 days, take the gas in the bottle and use the method of stable isotope ratio mass spectrometry to detect whether stable isotope-labeled nitrogen has been produced.

[0049] Nitrogen detection medium formula (g / L): 15 N stable isotope labeled NH 4 Cl 0.19, sodium hexahydrate 22.51, KH 2 PO 4 0.5, Na 2 HPO 4 12H 2 O 1.25, MgSO 4 ·7H 2 O 0.2, trace element sol...

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Abstract

The present invention provides an Alcaligenes sp. strain HO‑1 with a preservation number of CGMCC No. 16549, which is isolated from a semi-short-path nitrification reactor for treating pig farm wastewater. The strain has been deposited in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures on September 28, 2018, and the preservation number is CGMCC No.16549. Alcaligenes provided by the present invention can efficiently remove ammonia nitrogen, and can independently convert NH 4 + converted to gaseous nitrogen product (N 2 O, N 2 ). When treating ammonia-nitrogen-containing wastewater, it can realize the whole process of denitrification under a single strain and a single condition, which is more advantageous than the traditional biological denitrification process of aerobic nitrification-anaerobic denitrification in terms of operation and economic benefits. In addition, the strain has a wide range of C / N ratio, temperature, pH and dissolved oxygen adaptability, and can tolerate and remove high concentrations of ammonia nitrogen.

Description

technical field [0001] The invention belongs to the technical field of environmental microorganism denitrification bacteria. [0002] technical background [0003] With the progress of society and the rapid development of industry and agriculture, human activities such as aquaculture, livestock and poultry breeding, urban life, and agricultural fertilization and irrigation have brought excessive nitrogen and organic matter emissions to the environment, resulting in eutrophication of water bodies and derivatives. A series of environmental problems such as algal blooms and black and odorous water bodies are becoming more and more serious. Effective denitrification treatment of sewage to ensure discharge standards is of great significance for controlling nitrogen discharge and protecting and treating water environment. [0004] Biological nitrogen removal is currently considered to be the most economical and effective way to remove nitrogen from sewage. The traditional biologi...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C02F3/02C02F3/34C12R1/05C02F101/16
CPCC02F3/02C02F3/34C02F2101/16C12N1/205C12R2001/05Y02W10/10
Inventor 刘志培刘双江吴梦茹刘缨苗莉莉高喜燕彭积森
Owner INST OF MICROBIOLOGY - CHINESE ACAD OF SCI
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